Background and purpose:Ovarian cancer is associated with a high recurrence and mortality due to the existence of cancer stem cells. The purpose of this study was to investigate the effect of casticin (CAS) on the capability of self-renewal in ovarian cancer stem cell like cells (OCSLCs) derived from SKOV3 cell line. Methods:SKOV3 cell line cells were cultured in vitro, and OCSLCs were obtained and amplified through suspended culture with conditioned medium of the stem cells. The phosphorylation level of FoxO3a was analyzed using Western blot. The protein expression of FoxO3a was inhibited by FoxO3a speciifc siRNA transfection, and then the ratio of sphere-formation was detected. Results: Compared with parental cells, OCSLCs over-expressed phosphorylated FoxO3a (pFoxO3a) and had elevated ratio of sphere-formation [(3.1±0.3)% vs (34.8±6.8)%, P<0.05]. CAS significantly inhibited the capability of sphere-formation in OCSLCs and down-regulated the expression level of pFoxO3a. And the transfection of FoxO3a speciifc siRNA suppressed the protein expression of FoxO3a and attenuated the inhibitory effect of CAS on the sphere-formation of OCSLCs. Conclusion: Reduced expression level of pFoxO3a is involved in the effect that CAS inhibits sphere-formation of OCSLCs derived from SKOV3 cell line.

Objective To explore whether Forkhead O transcription factor-3a (FoxO3a) activity affects the capability of sphere-formation of ovarian cancer SKOV3 cell line.Methods Sphere-forming cells (SFCs) were obtained and amplified through suspended culture with conditioned medium of the stem cells in SKOV3 cell line.After SKOV3 cells were transfected with FoxO3a specific siRNA,the protein expressions of FoxO3a and Bmi1 and the ratio of sphere-formation were compared with Western blot and sphere-forming assay,respectively.Results Compared to parental cells,SFCs from SKOV3 cell line had higher ratio of sphere-formation and over-expressed Bmi1 and pFoxO3a.Transfection of FoxO3a specific siRNA down-regulated the protein expression of FoxO3a and upregulated expression of Bmi1 in SKOV3 cells,and enhanced the capability of sphere-formation.Conclusions Silence of FoxO3a leads to enhanced capability of sphere-formation in SKOV3 cell line.

Hoxa3-Pax1/Pax9-Eya1-Six1/4 regulatory pathway seems to be operating during forming the bilateral parathyroid/thymus common primordial in early embryonic development.The specification of the parathyroid domain in the parathyroid/thymus primordial is regulated through a Shh-Tbx1-Gcm2 pathway.Gcm2 also may play roles in later steps of parathyroid development,including CaSR and PTH gene expression.MafB and Gcm2 interact with each other and synergistically activate PTH transcription.Genetic basis and the etiology of some hypoparathyroid disorders in man are involved defects in transcription factors that include GCMB,GATA3,Tbxl,SOX3 and GNA11.This marker expression in thymus and parathyroid primordium includes HoxA3,Pax1,Eya1,and Six1;and expression of parathyroid cell-like cells includes Gcm2,CaSR,and PTH.These expressions may serve as markers of stem cell differentiation into parathyroid cell-like cells.

Objective To investigate the effects of different doses of alcohol on the synthesis of testosterone and the expression of androgen binding protein(ABP)mRNA in rat testis.Methods Forty male Wistar rats were randomly divided into 4 groups(10 rats each group)and received either distilled water(control group)or alcohol(alcohol-fed groups)for 5 months.Alcohol was administered by garage with a single daily dose : 5 g/kg(large dose group),2.5 g/kg(middle dose group)and 0.5 g/kg(small dose group).Testosterone content was measured by ELISA.mRNA levels of peripheral-type benzodiazepine receptors(PBR),PPARct and ABP were assayed by RT-PCR.Results Compared with control group:(1)ethanol feeding with daily doses of 5 g/kg,2.5 g/kg and 0.5 g/kg significantly decreased testosterone levels by 31.13%(P0.05)respectively,indicating that ethanol might impair testosterone synthesis;(2) mRNA levels of PBR were decreased in all three ethanol-treated groups(all P

Objective To detect the expression of interlenkin-22 (IL-22) and relative CD4+ T cell subsets in patients with ulcerative colitis (UC),and to explore their roles in the pathogenesis of UC.Methods Thirty-five adult UC patients were enrolled in this study and 35 healthy subjects were taken as control.Plasma IL-22 level was quantified by ELISA.The percentages of Th1,Th17 and Th22 cells in peripheral blood were determined by flow cytometry.The relationships of these results and disease activity were analyzed.Results Plasma IL-22 levels in UC patients [ (354.12±104.22 ) pg/ml ]were significantly higher than that of healthy controls ( P＜0.05 ),and the levels increased significantly in severely active patients.The percentages of Th17 cells in UC patients [ (2.36±0.94) ％ ]were elevated compared to healthy controls ( P＜0.05 ),and the percentages increased significantly in moderately active and severely active patients.The percentages of Th22 cells in UC patients [ (2.27±0.87 ) ％ ]were elevated compared to healthy controls (P±0.05),and the percentages increased significantly in severely active patients.The percentage of Th1 cells was not significantly different between UC patients and normal controls.ConclusionOur resuits demonstrate elevated IL-22 correlated to Th17 and Th22 cells may play an important role in the immunopathogenesis of UC.

BACKGROUND:Cytokine induced kil er cells therapy as an effective means of adoptive immunotherapy, becomes a new way to treat acute myeloid leukemia. But, the researches about sequential cytokine induced kil er cells therapy after autologous peripheral blood stem celltransplantation in acute myeloid leukemia patients are stil less, which deserve further research.
OBJECTIVE:To observe the clinical efficiency and safety of sequential cytokine induced kil er cells therapy after autologous peripheral blood stem celltransplantation in acute myeloid leukemia M2 patients.
METHODS:Total y 45 patients with low-or intermediate-risk acute myeloid leukemia M2 were recruited in this study. Among them, 19 patients received sequential cytokine induced kil er cells therapy after autologous peripheral blood stem celltransplantation and 26 patients only received autologous peripheral blood stem celltransplantation. The relapse rate, disease-free survival, and overal survival were compared between two groups, and safety of cytokine induced kil er cells therapy was observed.
RESULTS AND CONCLUSION:(1) Compared with the patients only receiving autologous peripheral blood stem celltransplantation, the relapse rate was lower (21.05%vs. 38.46%;P<0.05), and elevated percentages of the disease-free survival and overal survival were observed in the patients receiving sequential cytokine induced kil er cells therapy after autologous peripheral blood stem celltransplantation (P<0.05). (2) The 19 patients who received sequential cytokine induced kil er cells therapy after autologous peripheral blood stem celltransplantation al completed the treatment scheme successful y. Only four patients appeared to have chil s and fever, and no more side effects were observed. These findings suggested that the sequential cytokine induced kil er cells therapy after autologous peripheral blood stem celltransplantation can improve the disease-free survival and overal survival of low-or intermediate-risk acute myeloid leukemia M2 patients without remarkable side effects, which is a safe, effective and feasible way for the treatment of acute myeloid leukemia M2.

Objectives To discuss the clinical characteristic, cause and measures to prevention and control of nosocomial infection in a neonatal intensive care unit (NICU). Methods Retrospectively analyzed an nosocomial infection outbreak of Klebsiella pneumoniae in NICU. Results From Sept. 3, 2010 to Oct. 3, 2010, there were 7 cases of hospital infection in 12 cases of sputum cultured Klebsiella Pneumoniae. The gestational age (GA) of 7 hospital infection cases was 28.5±2.6 week. The birth weight of infection cases was 941.4±309.8 g. The onset of infection was at 31.7±12.8 d of hospitalization. The nosocomial incidence was 2.41%in the hospital, which was 5.79%in preterm infants, 50.00%in GA<28w infants, and 42.86%in extremely low birth weight infant (ELBW). All sputum culture results were displayed as multi-drug resistant of Klebsiella pneumoniae, penicillin and third-generation cephalosporin antibiotic resistance rate of 75%to 100%. The resistance rates to penicillin and cephem antibiotics were 75% -100%, carbapenems was 58.3%, piperacillin/tazobactam was 25.0%. All nosocomial patients were cured. Conclusions GA<28w and ELBW infants are at increased risk of nosocomial infection in NICU. The emergence of carbapenems resistant Klebsiella Pneumoniae has been increasing with the widespread use of carbapenems. Hospital infection can be controlled by standardized medical behavior, which can decline the nosocomial infection incidence and mortality of preterm infants in NICU.

Objective To explore the expression of HCK gene during the cardiomyocyte differentiation of mouse embryonic stem cells and analyze the role of HCK gene in maintenance of pluripotency of embryonic stem cells.Methods Mouse embryonic stem cells were cultured,then induced to differentiate into cardiomyocytes.Total RNAs were isolated from mouse embryonic stem cells in the differentiation days:0 day(D0),the second day(D2),the fourth day(D4),the sixth day(D6),the eighth day(D8),respectively.The levels of HCK mRNAs were assessed by the method of semi-quantitive reverse transcriptase-polymerase chain reaction(RT-PCR).In the meanwhile,Total proteins were also isolated from mouse embryonic stem cells in the differentiation D0,D2,D4,D6,D8,and the levels of HCK proteins were evaluated by Western-blot.Results HCK mRNAs could be detected in the mouse embryonic stem cells in D0 and D2,however,they were undetectable from D4 to D8.The expression of HCK mRNAs was rapidly down-regulated during cardiomyocyte differentiation of mouse embryonic stem cells.Expression of HCK proteins,which coincided with HCK mRNAs,down-regulated during differentiation and couldn't be detected in D4.Conclusions With the cardiomyocyte differentiation of mouse embryonic stem cells,the expression of HCK in the levels of mRNA and proteins are sharply down-regulated;HCK may play an important role in maintaining the pluripotency of embryonic stem cell.

The aim of this study is to investigate the anti-inflammatory effect of the adenosine derivative N6-(3-hydroxylaniline) adenosine (WS070117M1) on cigarette smoke plus LPS (lipopolysaccharide)-induced chronic obstructive pulmonary disease (COPD) in mice and its mechanism. COPD model was established by exposing male BALB/c mice to cigarette smoke and challenged with LPS inhalation. Supernatants of bronchoalveolar lavage fluid (BALF) were harvested and IL-1β, IL-6, IL-8 and TGF-β1 levels were measured by ELISA (enzyme-linked immunesorbent assay). The number of total white blood cells and neutrophils in bronchoalveolar lavage fluid was counted separately. Lung tissue was stained with Mayer 's hematoxylin and eosin for histopathologic examination. pAMPKa protein expression and distribution of lung tissue were analyzed by immunohistochemistry method. In vitro, levels of AMPKα phosphorylation in phorbol-12- myristate-13-acetate (PMA) differentiated THP-1 cells was detected by immunohistochemistry, IL-8 level in supernatants of cigarette smoke condensate stimulating PMA differentiated THP-1 cells was measured by ELISA. The results showed that WS070117M1 treatment significantly activated AMPKa in the lung tissue. It also resulted in down regulation of IL-1β, IL-6, IL-8 and TGF-β1 levels in bronchoalveolar lavage fluid and IL-8 level in cigarette smoke condensate stimulating PMA differentiated THP-1 cells. In addition, WS070117M1 could inhibit the recruitment of total white blood cells and neutrophils. These results suggest that WS070117M1 may alleviate the airway inflammation by activating AMPK in the lung tissue.
AMP-Activated Protein Kinases ; metabolism ; Adenosine ; analogs & derivatives ; Animals ; Bronchoalveolar Lavage Fluid ; Cell Line, Tumor ; Disease Models, Animal ; Humans ; Inflammation ; drug therapy ; Interleukin-1beta ; metabolism ; Interleukin-6 ; metabolism ; Interleukin-8 ; metabolism ; Leukocyte Count ; Lipopolysaccharides ; Male ; Mice ; Mice, Inbred BALB C ; Neutrophils ; cytology ; Pulmonary Disease, Chronic Obstructive ; drug therapy ; Smoke ; adverse effects ; Tobacco ; Transforming Growth Factor beta1 ; metabolism

A cross-sectional survey with multiple-stage and random sampling was performed among middle and elderly aged permanent inhabitants in Wuhan area.The prevalences of metabolic syndrome,diabetes mellitus, impaired glucose tolerance,hypertension and obesity were 12.2%,11.8%,10.3%,31.9% and 48.0% respectively.